Simultaneous 3D cellular positioning and apical dendritic morphology of transgenic fluorescent mouse CA3 hippocampal pyramidal neurons.
Simultaneous 3D cellular positioning and apical dendritic morphology of transgenic fluorescent mouse CA3 hippocampal pyramidal neurons.
复制标题
转基因荧光小鼠 CA3 海马锥体神经元的同步 3D 细胞定位和顶端树突形态。
DOI：
10.1016/j.jneumeth.2023.109823
复制
发表时间：
2023
影响因子：
3
通讯作者：
Vidal,GeorgeS
中科院分区：
文献类型：
--
作者：
Handwerk,ChristopherJ;Bland,KatherineM;Denzler,CollinJ;Kalinowski,AnnaR;Brett,CooperA;Swinehart,BrianD;Rodriguez,HildaV;Cook,HollynN;Vinson,ElizabethC;Florenz,MadisonE;Vidal,GeorgeS
研究背景海马CA 3区锥体神经元的树突形态多样，结构和功能也不尽相同。然而，很少有结构研究捕捉到精确的三维躯体位置和三维树突状形态的CA 3锥体neurons.New methodHere，我们提出了一个简单的方法来重建顶端树突状形态的CA 3锥体神经元使用转基因fluorescentThy 1-GFP-M线。该方法同时跟踪海马内重建神经元的背腹、切线和径向位置。它是专门设计用于与转基因荧光小鼠线，这是常用的神经元形态和development.ResultsWe遗传研究中的地形和形态数据是如何捕获的转基因荧光小鼠CA 3锥体neurons.Comparison与现有的methodsThere是没有必要选择和标记CA 3锥体神经元与转基因fluorescentThy 1-GFP-M线。通过采取横向（而不是冠状）连续切片，我们保留了精细的背腹，切线和径向体定位的三维重建神经元。由于PCP 4免疫组化能很好地确定CA 2的位置，因此我们在此使用该技术来提高沿CA 3沿着切向定位的精确性。结论我们建立了一种同时收集转基因荧光小鼠海马锥体神经元精确的体细胞定位和三维形态学数据的方法。这种荧光方法应与许多其他转基因荧光报告线和免疫组化方法兼容，便于捕获的地形和形态学数据，从各种各样的遗传实验在小鼠海马。
BackgroundPyramidal neurons throughout hippocampal CA3 are diverse in their dendritic morphology, and CA3 is not homogenous in its structure or function. Nonetheless, few structural studies have captured the precise 3D somatic position and the 3D dendritic morphology of CA3 pyramidal neurons simultaneously.New methodHere, we present a simple approach to reconstruct the apical dendritic morphology of CA3 pyramidal neurons using the transgenic fluorescentThy1-GFP-M line. The approach simultaneously tracks the dorsoventral, tangential, and radial positions of reconstructed neurons within the hippocampus. It is especially designed for use with transgenic fluorescent mouse lines, which are commonly used in genetic studies of neuronal morphology and development.ResultsWe demonstrate how topographic and morphological data are captured from transgenic fluorescent mouse CA3 pyramidal neurons.Comparison with existing methodsThere is no need to select and label CA3 pyramidal neurons with the transgenic fluorescentThy1-GFP-M line. By taking transverse (not coronal) serial sections, we preserve fine dorsoventral, tangential, and radial somatic positioning of 3D-reconstructed neurons. Because CA2 is well defined by PCP4 immunohistochemistry, we use that technique here to to increase precision in defining tangential position along CA3.ConclusionsWe developed a method for simultaneously collecting precise somatic positioning as well as 3D morphological data among transgenic fluorescent mouse hippocampal pyramidal neurons. This fluorescent method should be compatible with many other transgenic fluorescent reporter lines and immunohistochemical methods, facilitating the capture of topographic and morphological data from a wide variety of genetic experiments in mouse hippocampus.